Recombinant SARS-CoV-2 Y453F Spike RBD His-tag Protein, CF

Catalog #: 10744-CV Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
10744-CV-100

Recombinant SARS-CoV-2 Y453F Spike RBD His-tag Protein Binding Activity

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Product Details

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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant SARS-CoV-2 Y453F Spike RBD His-tag Protein, CF Summary

Product Specifications

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<0.10 EU per 1 μg of the protein by the LAL method.

Activity

Measured by its binding ability in a functional ELISA with Recombinant Human ACE-2 His-tag (Catalog # 933-ZN).

Source

Human embryonic kidney cell, HEK293-derived sars-cov-2 Spike RBD protein
Arg319-Phe541 (Tyr453Phe), with a C-terminal 6-His tag

Accession #

YP_009724390.1

N-terminal Sequence
Analysis

Arg319

Predicted Molecular Mass

26 kDa

SDS-PAGE

31-37 kDa, under reducing conditions

Product Datasheets

10744-CV

Product Datasheet

COA

SDS

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

10744-CV

Formulation	Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
Reconstitution	Reconstitute at 500 μg/mL in PBS.
Shipping	The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage:	Use a manual defrost freezer and avoid repeated freeze-thaw cycles. 12 months from date of receipt, -20 to -70 °C as supplied. 1 month, 2 to 8 °C under sterile conditions after reconstitution. 3 months, -20 to -70 °C under sterile conditions after reconstitution.

Scientific Data

Binding Activity

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Recombinant SARS-CoV-2 Y453F Spike RBD His-tag (Catalog # 10744-CV) binds Recombinant Human ACE-2 His-tag (933-ZN) in a functional ELISA.

SDS-PAGE

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2 μg/lane of Recombinant SARS-CoV-2 Y453F Spike RBD His-tag (Catalog # 10744-CV) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 31-37 kDa and 28-35 kDa, respectively.

Surface Plasmon Resonance (SPR)

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Recombinant SARS-CoV-2 Spike RBD Y453F His-tag was immobilized on a Biacore Sensor Chip CM5, and binding to recombinant human ACE-2 (933-ZN) was measured at a concentration range between 0.18 nM and 94.3 nM. The double-referenced sensorgram was fit to a 1:1 binding model to determine the binding kinetics and affinity, with an affinity constant of KD=2.430 nM.

Reconstitution Calculator

Background: Spike RBD

SARS-CoV-2, which caused the global pandemic coronavirus disease 2019 (Covid-19), belongs to a family of viruses known as coronaviruses that also include MERS and SARS-CoV-1. Coronaviruses are commonly comprised of four structural proteins: Spike protein (S), Envelope protein (E), Membrane protein (M) and Nucleocapsid protein (N) (1). The SARS-CoV-2 S protein is a glycoprotein that mediates membrane fusion and viral entry. The S protein is homotrimeric, with each ~180-kDa monomer consisting of two subunits, S1 and S2 (2). In SARS-CoV-2, as with most coronaviruses, proteolytic cleavage of the S protein into S1 and S2 subunits is required for activation. The S1 subunit is focused on attachment of the protein to the host receptor while the S2 subunit is involved with cell fusion (3-5). A metallopeptidase, angiotensin-converting enzyme 2 (ACE-2), has been identified as a functional receptor for SARS-CoV-2 through interaction with a receptor binding domain (RBD) located at the C-terminus of S1 subunit (6,7). The RBD of SARS-CoV-2 shares 73% amino acid (aa) identity with the RBD of the SARS-CoV-1, but only 22% aa identity with the RBD of MERS. A SARS-CoV-2 variant carrying the aa substitution Y453N in the RBD was first identified in farmed minks (8). The Spike RBD of this new virus variant was reported to show increased affinity to ACE-2, but whether the Y453N mutation in RBD would decrease the efficacy of vaccine‑induced immunity is still under investigation (9, 10).

References

Wu, F. et al. (2020) Nature 579:265.
Tortorici, M.A. and D. Veesler (2019). Adv. Virus Res. 105:93.
Bosch, B.J. et al. (2003). J. Virol. 77:8801.
Belouzard, S. et al. (2009) Proc. Natl. Acad. Sci. 106:5871.
Millet, J.K. and G.R. Whittaker (2015) Virus Res. 202:120.
Li, W. et al. (2003) Nature 426:450.
Wong, S.K. et al. (2004) J. Biol. Chem. 279:3197.
Oreshkova, N. et al. (2020) Euro. Surveill. 25:2001005.
Bayarri-Olmos, R. et al. (2020) bioRxiv https://doi.org/10.1101/2021.01.29.428834.
Hayashi, T. et al. (2020) bioRxiv https://doi.org/10.1101/2020.11.27.401893.